122nd General Meeting of the KCS

Type Poster Presentation
Area Life Chemistry
Room No. Grand Ballroom
Time 10월 19일 (금요일) 11:00~12:30
Code LIFE.P-415
Subject Application of the Red Fluorescent Protein as an Indicator of Recombinant Protein Production in Escherichia coli
Authors Yoonjin Um, Young Kee Chae*
Department of Chemistry, Sejong University, Korea
Abstract The discovery of green fluorescent protein has brought a new era in cell biology by enabling scientists to monitor various cellular processes in living systems in real time via optical microscopy by fusing the fluorophore moiety to many proteins and enzymes of interest. Researchers have developed a broad spectrum of fluorescent proteins, covering the entire visible light range from blue to green to yellow to red colors. We have employed the red fluorescent protein (mCherry) to provide the visualization of the target gene expression in Escherichia coli. The mCherry gene was tagged right next to the maltose-binding protein (MBP) which was widely used as a fusion partner to provide high yield and solubility. The target gene was inserted behind the His-Tag, and the tagged protein was designed to be cleaved off of the MBP-mCherry inside the cell by the constitutively expressed TVMV protease. The red color became clearly visible after the induction signal was turned on, showing that the expression of the MBP-mCherry-target gene had started and went well. The target protein was successfully purified by by the Ni-NTA column. This work has been supported by Science Research Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Education (NRF-2017R1D1A1A02017545). References 1. Hui, C.-Y., Guo, Y., Zhang, W., & Huang, X.-Q. (2018). Rapid monitoring of the target protein expression with a fluorescent signal based on a dicistronic construct in Escherichia coli. AMB Express, 8, 81. http://doi.org/10.1186/s13568-018-0612-5 2. Su, W. W. (2005). Fluorescent proteins as tools to aid protein production. Microbial Cell Factories, 4, 12. http://doi.org/10.1186/1475-2859-4-12 3. Using green and red fluorescent proteins to teach protein expression, purification, and crystallization. Biochem Mol Biol Educ (2008) 36(1), 43. http://doi.org/10.1002/bmb.117
E-mail ykchae@sejong.ac.kr